Figure 6From: Promoter methylation inhibits BRD7 expression in human nasopharyngeal carcinoma cells In vitro cytosine methylation of BRD7 promoter silence its activity in NPC cells. (A) Detection of the methylation effects of Met-pGL3-404/+46 and Met-pGL3-404/+46/GFP by restriction enzyme cutting of HpaII. pGL3-404/+46 was used as a control. (B) Detection of the luciferase activity of methylated BRD7 promoter construct Met-pGL3/-404,+46 in COS7, BHK-21, HNE1, CNE1, 6–10B, 5–8F, SW480, and Hela cells. Luciferase activity in COS7 and 5–8F cells is represented by black and gray histograms, respectively. All of the constructs were cotransfected with the SV40 β-galactosidase vector for normalizing transfection efficiency. Data are the means ± S.D. of three independent experiments. (C) Detection of the luciferase activity of modified GFP reporter construct Met-pGL3/-404,+46/EGFP in 5–8F cells. The full-length modified promoter construct pGL3/-404,+46/EGFP was used as a positive control. 38 h after transfection, the signal of EGFP fluorescence driven by promoter fragment -404/+46 or -266/-212 was observed by using an AX-80 analytical microscope system (Olympus, Tokyo, Japan).Back to article page